Vol 8, No 2 (2013)

Glioblastoma multiforme is a malignant primary brain tumor with a very poor prognosis. Neural stem and progenitor cells of the adult brain, as well as other types of stem cells are considered by carcinogenesis researchers the most likely source of malignant gliomas. This is evidenced by common genes regulating key processes of life, uniform proteomic profiles and identical immunophenotypical cell surface markers. These cells are highly proliferative, multipotent, able to independently migrate to the damaged area and have extensive replicative potential. However, antitumor properties of the stem cells (SCs) are also confirmed. The bodies of adult mammals and humans have genetically fixed mechanisms of control over their populations and multiple levels of antitumor protection. So far, the role of autologous SCs in a tumor patient is not clear. On the one hand, they fail to fulfill their anti-tumor and regulatory functions, and instead of organizing anti-tumor response become one of the key elements of carcinogenesis contributing to the development of neoplastic tumor vascular network and modulating the processes of neurogenesis, which is the main source of its pathological reinnervation, and therefore, a pain. On the other hand, autologous stem cells trigger powerful proliferative processes in the tumor tissue becoming the driving force of neoplastic growth. Apparently, due to the interaction of tumor cells with autologous stem cells, general and local interstitial patterns of autoregulation and sanogenesis are disturbed making tumor growth possible in principle. From this perspective, the technology that directly affects the population of cancer stem cells seems the most promising.

The choice object for cell therapy of degenerative changes of the intervertebral discs is a cryopreserved multipotent mesenchymal stromal cells (MMSC). The research aim was to evaluate a therapeutic potential of cryopreserved allogenic marrow-derived MMSC in this pathology. Rats with modeled compressive degenerative damage of the intervertebral disc CcVI-VII were introduced by 0.5x10 B native or cryopreserved as well as PKH-26-labeled cells on a collagen sponge in the defect area. Animals with spontaneous recovery and administration of saline were served as a control. Histomorphometric study and fluorescent microscopy of intervertebral disc sections was performed on the 30th day after treatment. There was extremely low regenerative potential in the control groups of animals. Histologically after cell therapy, there was tendency to repair of cracks, fissures, collagen fiber fragmentations of the fibrous ring, which was more pronounced in the case of the native MMSC culture application. The cryopreserved MMSC administration was accompanied by increasing in the fibrochondrocyte number of the dorsal annulus per unit area of the intervertebral disc CcVI-VII slice at 1.25 times relative to the model values. At the same time the fibrous ring height increased by 12.5±3.3%, and densitometric index of cartilaginous tissue — by 64±5.7% relative to the model values. The luminescence in the red range of the spectrum in the form of drops and conglomerates, which was localized in outer parts of fibrous ring, was detected by fluorescent microscopy of intervertebral disc cryostat sections on the 30 day after introduction of labeled native or cryopreserved MMSC, that indicated the safety and partial migration of transplanted and/ or their daughter cells in the intervertebral disc CcVI-VII. Our data showed a stimulating effect of native and cryopreserved marrow-derived MMSC suspension in degenerative intervertebral disc damages.

Generation of human cardiomyocytes in vitro is important for basic science and for regenerative medicine. There are a lot of published protocols describing the cardiomyocytes obtaining with different efficacy. However in practice, a researcher is faced with the fact that published methods have to be adapted to a particular laboratory for specific tasks. The aim of this study was to compare three methods for generation of human cardiomyocytes in vitro: 1) from human adult heart sample by purifying myocardial progenitor cells and their subsequent differentiation into cardiomyocyte direction; 2) from human adipose tissue multipotent mesenchimal stromal cells (AT-MSC); 3) from human dermal fibroblasts using reprogramming technology and subsequent differentiation to cardiomyocytes. We have shown relatively low efficiency of cardiac progenitor cells and AT-MSC capacity to give rise to cardiomyocyte lineage. The effectiveness of cardiomyocytes differentiation from dermal fibroblasts by reprogramming technology was significantly higher. The advantages, disadvantages and perspectives of each method are discussed.

Solid-pseudopapillary tumor (SPPO) is a rare tumor of a pancreas with unknown histogenesis. In this regards some theories about its development are discussed including genesis from so called cancer stem cells, which can be derivative of embryonic ovaries primordiums. We studied 11 SPPO and 5 ovarian granulosa cell tumors (GCT) adult type by electron microscopy. A comparative ultrastructural study identified similar features in GCT and SPPO: the presence of two cells types, one of which with characteristic nuclei with deep invaginations nukleolemmy as longitudinal furrows, intercellular contacts in the form of desmosomes, in the tissues quite large number of capillaries in the active state were detected. Among all ovarian tumors developing from cells of sex cord stroma SPPO is the most close to GCT. They are related not only due to morphological similarity, but the biological potential, the clinical course and disease prognosis. Our studies confirm the theory of embryonic migration of ovaries primordiums cells with their subsequent contribution to SPPO histogenesis.

Transplantation of allogenic hematopoietic stem cells used for treatment of oncological and immune diseases during the late 1970s. Despite of that the matched donor search is the main problem in this field as before. To resolve this problem the bone marrow donor registries were established around the world. In 2010 Bone marrow donor registry (BMDR) was established in Clinical Centre of Cell Technologies. BMDR includes adult donors from many regions of Russia and cord blood units (CBUs) from Samara cord blood bank. At the present day, there are more than 5000 CBUs and 5000 donors. During the work of the registry 44 CBUs were handed to Russian and international transplant centers. In according with received results, the survival rate was about 65%. In relation with increase of oncohematological diseases in Russia the development of national bone marrow donor registry is extremely important.

We performed proteome mapping, cataloguing and bioinformation analysis of protein lysates of human neural (CD133+) stem cells (NSC) isolated from olfactory sheath of a nose, multipotent mesenchymal (CD29+, CD44+, CD73+, CD90+, СD34 -) stromal cells (MMSC) isolated from human bone marrow and tumor (CD133+) stem cells (TSC) isolated from the human U87 glioblastoma cell line. We identified 1664 proteins in the examined lysates of stem cells (SC), 1052 (63.2%) of which are identical in NSC and TSC and 607 proteins (36.47%) are identical in MMSC and TSC. Other proteins in U87 glioblastoma TSC are oncospecific or carcinogenesis associated. The biological processes, molecular functions, cell localization and protein signal pathways of the proteins available in all three proteomes were annotated by PubMed, PANTHER, Gene Ontology and KEGG databases. It was shown that gliomaspheres of U87 glioblastoma had only 10 intracellular pathways of signal transduction (IPST) that were not modified by neoplastic process, but only two of them (integrin and focal adhesion pathways) were accessible for regulatory action on gene candidates in TSC nucleus. Carcinogenesis free membrane proteins, intracellular pathways of signal transduction and genes expressing proteins of these pathways in U87 glioblastoma TSC can be viewed as main targets for regulatory effect on TSC. We offer a novel concept of proteome-based complex therapy of tumors.